U.S. patent application number 14/016060 was filed with the patent office on 2014-03-06 for method of tracking specific cells in vivo.
This patent application is currently assigned to ACADEMIA SINICA. The applicant listed for this patent is ACADEMIA SINICA. Invention is credited to Chia-Chi Chien, Yeu-Kuang Hwu, Cheng-Liang Wang.
Application Number | 20140066762 14/016060 |
Document ID | / |
Family ID | 50188437 |
Filed Date | 2014-03-06 |
United States Patent
Application |
20140066762 |
Kind Code |
A1 |
Hwu; Yeu-Kuang ; et
al. |
March 6, 2014 |
METHOD OF TRACKING SPECIFIC CELLS IN VIVO
Abstract
A method of tracking specific cells in vivo is disclosed. The
method of the disclosure includes: providing fluorescent
nanoparticles suitable for targeting of specific cells;
administering the fluorescent nanoparticles to a subject; providing
an X-ray source to irradiate the subject; and determining the
distribution and growth of the specific cells by the fluorescent
images from the fluorescent nanoparticles and X-ray images of the
subject irradiated by the X-ray source.
Inventors: |
Hwu; Yeu-Kuang; (Taipei,
TW) ; Chien; Chia-Chi; (Taipei, TW) ; Wang;
Cheng-Liang; (Taipei, TW) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ACADEMIA SINICA |
Taipei |
|
TW |
|
|
Assignee: |
ACADEMIA SINICA
Taipei
TW
|
Family ID: |
50188437 |
Appl. No.: |
14/016060 |
Filed: |
August 31, 2013 |
Current U.S.
Class: |
600/431 ;
977/905 |
Current CPC
Class: |
B82Y 5/00 20130101; A61B
6/508 20130101; B82Y 15/00 20130101; A61B 6/481 20130101; A61K
49/0065 20130101; A61B 6/485 20130101 |
Class at
Publication: |
600/431 ;
977/905 |
International
Class: |
A61B 6/00 20060101
A61B006/00 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 31, 2012 |
TW |
101131694 |
Claims
1. A method of tracking specific cells in vivo, comprising:
providing a plurality of fluorescent nanoparticles suitable for
targeting specific cells; administering the plurality of
fluorescent nanoparticles to a subject; providing an X-ray source
to irradiate the subject; and determining the growth and
distribution of the specific cells by fluorescent images of the
plurality of fluorescent nanoparticles and the X-ray images from
the subject irradiated by the X-ray source and observing the growth
and distribution of a capillary in vivo by the X-ray images of the
plurality of fluorescent nanoparticles.
2. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the plurality of fluorescent nanoparticles
comprise a nanoparticle including Gd.sub.2O.sub.3,
Y.sub.3Al.sub.5O.sub.12, Y.sub.2SiO.sub.5, ZnO,
BaMgAl.sub.14O.sub.23, Ti.sub.2O.sub.3, Zn.sub.2SiO.sub.2,
Cn.sub.2SiO.sub.4, BaSiO.sub.4, or (Y,Gd)BO.sub.3.
3. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the plurality of fluorescent nanoparticles have a
diameter between about 1 nm and 100 .mu.m.
4. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the X-ray source comprises a synchrotron radiation
X-ray source, a medical X-ray source, or a laboratory X-ray
source.
5. The method of tracking specific cells in vivo as claimed in
claim 4, wherein a photon energy of the X-ray source is between
about 4 keV and 20 MeV.
6. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the absorbed dose of the X-ray source in the
subject is less than about 100 Gy.
7. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the absorbed dose of the X-ray in the subject is
between about 1 Gy and 30 Gy.
8. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the irradiation time of the X-ray source to the
subject is less than about 30 minutes.
9. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the irradiation time of the X-ray to the subject
is between about 100 milliseconds and 5 minutes.
10. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the subject comprises humans, mammals, birds,
amphibians, reptiles, fish, insects, or other appropriate
multicellular animals.
11. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the specific cells comprise tumor cells, stem
cells, blood cells, tissue cells, or other appropriate somatic
cells.
12. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the X-ray image comprises vascular development or
cell targeting.
13. The method of tracking specific cells in vivo as claimed in
claim 1, wherein the effective penetration depth of the subject
irradiated by the X-ray source is about 30 cm from the surface to
the deep tissue.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority of Taiwan Patent
Application No. 101131694 filed on Aug. 31, 2012, the entirety of
which is incorporated by reference herein.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates to a method of tracking cells
in vivo, and in particular, relates to a method of tracking
specific cells in vivo using fluorescent nanoparticles.
[0004] 2. Description of the Related Art
[0005] In recent years, a variety of imaging methods have been
developed for biological and medical applications. Especially,
molecular imaging has been widely used to trace specific cells as a
diagnosis tool for tumor cells treatments. There are many studies
related to the application of fluorescent nanoparticles with
luminescent properties to fluorescence imaging. In vivo small
animal imaging, allows fluorescent images from fluorescent
particles to be observed by a high-sensitivity camera; however,
applications are limited as penetration of photons in tissue in
vivo are often inadequate. Fluorescent particles that emit the
near-infrared region (NIR) with high penetration are used for
enhancement. Currently, small molecules of indocarbocyanine dyes
are mainly used.
[0006] As the applications of fluorescent probes and fluorescent
reporters become wider, fluorescent imaging has become an important
analysis tool between basal and clinical researches. Although
traditional small molecule near-infrared region (NIR) dyes are
still used, development of fluorescent organic nanoparticles,
fluorescent biological nanoparticles, and fluorescent inorganic
nanoparticles for in vivo fluorescent imaging allow for the
development of many powerful new tools for biological medical
applications. Nanoparticles, as a platform, can be built up with
multi-functional probes to be applied in multimodality imaging.
[0007] In view of this, a novel imaging method which can be
combined with high-resolution and long life cycle fluorescent
imaging, molecular targeting techniques, and X-ray imaging to
function as a biomedical diagnosis tool is needed.
BRIEF SUMMARY OF THE INVENTION
[0008] A detailed description is given in the following embodiments
with reference to the accompanying drawings.
[0009] In one embodiment, the present disclosure provides a method
of tracking specific cells in vivo, which includes providing
fluorescent nanoparticles suitable for targeting specific cells,
administering the fluorescent nanoparticles to a subject, providing
an X-ray source to irradiate the subject, and determining the
growth and distribution of the specific cells by fluorescent images
of the fluorescent nanoparticles and X-ray images from to the
subject irradiated by the X-ray source and observing the growth and
distribution of a capillary in vivo by X-ray images of the
fluorescent nanoparticles.
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] The present invention can be more fully understood by
reading the subsequent detailed description and examples with
references made to the accompanying drawings, wherein:
[0011] FIG. 1 is an X-ray image of tumor angiogenesis using
Gd.sub.2O.sub.3 as a contrast agent.
DETAILED DESCRIPTION OF THE INVENTION
[0012] The following description is of the best-contemplated mode
of carrying out the invention. This description is made for the
purpose of illustrating the general principles of the invention and
should not be taken in a limiting sense. The scope of the invention
is best determined by reference to the appended claims.
[0013] The present disclosure provides a method of tracking
specific cells in vivo, which includes administering fluorescent
nanoparticles suitable for targeting specific cells to a subject,
wherein the fluorescent nanoparticles are capable of specifically
targeting specific cells in vivo and being a contrast agent at the
same time. Next, a high-coherent X-ray source is used to irradiate
the subject, wherein the high-coherent X-ray source can stimulate
the fluorescent nanoparticles in the subject to emit fluorescence.
In some embodiments, the wavelength of fluorescence emitted by the
fluorescent nanoparticles may be 450-900 nm. Then, the growth and
distribution of the specific cells are determined by fluorescent
images of the fluorescent nanoparticles in the subject and the
X-ray images from the subject irradiated by the X-ray source.
[0014] In one embodiment, the fluorescent nanoparticles may be
Gd-based nanoparticles, such as Gd.sub.2O.sub.3, or may include:
Y.sub.3Al.sub.5O.sub.12, Y.sub.2SiO.sub.5, ZnO,
BaMgAl.sub.14O.sub.23, Ti.sub.2O.sub.3, Zn.sub.2SiO.sub.2,
Cn.sub.2SiO.sub.4, BaSiO.sub.4, or (Y, Gd)BO.sub.3 nanoparticles,
which may not only be stimulated by the X-ray source to emit
fluorescence, but also function as biomedical probes to facilitate
tracking of the distribution of the tumor cells in vivo. The above
characteristics may be applied on photodynamic therapy as a
powerful clinical diagnosis work. To achieve the purpose of
tracking tumor cells or imaging tumor vasculature, the fluorescent
nanoparticles may have a diameter from between about 1 nm and 100
.mu.m to show the location of tumors.
[0015] In one embodiment, the X-ray source used to trace tumor
cells in vivo may include a synchrotron radiation X-ray source, a
medical X-ray source, or a laboratory X-ray source. In one
embodiment, the X-ray source may have an intensity of about 4
keV-20 MeV. The absorbed dose of the X-ray source in the subject is
less than about 4 Gy, preferably between about 0.1 and 1 Gy.
[0016] Due to the high coherent X-ray source (4 keV-20 MeV),
photons can penetrate the body, the fluorescent nanoparticles
administrated in the subject and marked on the tumor cells to emit
fluorescence can be efficiently stimulated and the autofluorescence
background to the fluorescent images can be reduced. In addition,
when the dosage of the X-ray accumulated to a certain amount, the
irradiation time of the X-ray source to the subject may be less
than about 1 millisecond, preferably less than about 100
milliseconds. The effective penetration depth of the subject
irradiated by the X-ray source may be about 30 cm from the surface
to the deep tissue. Since the high-energy X-ray source adopted in
the present disclosure has a high penetration ability in vivo,
tumor cells in vivo may be monitored immediately by fluorescent and
X-ray images of the present disclosure, instead of having to
perform sample slicing from living subjects as conventional medical
imaging requires.
[0017] The present disclosure is suitable for tracking any kinds of
somatic cells in a subject, such as tumor cells, wherein the
subject may include humans, mammals, birds, amphibians, reptiles,
fish, insects, and/or other appropriate multicellular animals. In
one embodiment, by combining X-ray images, such as vascular
development, cell calibration, a combination thereof, or
fluorescence images, the growth and distribution of the traced
cells may be determined.
[0018] Further, the applications of the present disclosure on
photodynamic therapy (PDT), including using the high-energy X-ray
source of the present disclosure, are capable of penetrating into
deep living tissues, efficiently stimulating the photo-sensitive
drugs swallowed? by tumor cells and overcoming the traditional
inadequate penetration of light source in living tissues. The
fluorescent nanoparticles of the present disclosure may be an
excellent vector for photo-sensitive drugs, and the fluorescent
nanoparticles, such as Gd-based nanoparticles, may be applied on
photodynamic therapy (PDT) to treat cancer after being stimulated
by the X-ray source. Therefore, a real-time diagnosis and treatment
of cancer may be achieved by combining the fluorescent and X-ray
imaging systems of the present disclosure and photodynamic therapy
(PDT).
[0019] In summary, in comparison to conventional biomedical
imaging, the present disclosure has the following advantages: (1)
reduces the interference of the autofluorescence background to the
fluorescent images; and (2) provides immediate monitoring of tumor
cells in living tissues by fluorescent and X-ray imaging of the
present disclosure, instead of having to perform sample slicing
from living subjects as conventional medical imaging requires.
EXAMPLES
Example 1
X-ray Images of Tumor Vessel Proliferation In Vivo
[0020] The mice used in this example were BALB/c mice (purchased
from National Laboratory Animal Center, Taiwan) fed by the Academia
Sinica Institutional Animal Care and Committee (AS IACUC). All mice
were housed in individual cages (five per cage) and kept at
24.+-.2.degree. C. with a humidity of 40%-70% and a 12-hour
light/dark cycle.
[0021] 4-5 week old mice were anesthetize by intramuscular
injection of 10 .mu.l of Zoletil 50 (50 mg/kg; Virbac Laboratories,
Carros, France), and PE-08 was cathetered in the mice (about 20-25
g of weight). Then, 200 .mu.l, 10 mg/mL of a contrast agent with
Gd.sub.2O.sub.3 was injected from the femoral artery into the
late-stage tumor (16-day) of the mice through the above PE-08
catheters (BB31695, Scientific Commodities, Inc.: 0.2 mm, O.D.:
0.36 mm), wherein the injection velocity of each group of a
contrast agent was 1 .mu.l/s. During the imaging process, mice were
anesthetized under 1% isoflurene in oxygen. X-ray images were taken
after 1 minute starting from the injection of the contrast agent
from the femoral artery into the mice, and the exposure time was
100 milliseconds. FIG. 1 shows an X-ray image of tumor angiogenesis
in vivo using Gd.sub.2O.sub.3 as a contrast agent, and the
distribution of a capillary was clearly revealed by the contrast
agent.
Example 2
Fluorescent Images In Vivo
[0022] The mice used in this example were BALB/c mice (purchased
from National Laboratory Animal Center, Taiwan) fed by the Academia
Sinica Institutional Animal Care and Committee (AS IACUC). All mice
were housed in individual cages (five per cage) and kept at
24.+-.2.degree. C. with a humidity of 40%-70% and a 12-hour
light/dark cycle.
[0023] 4-5 week old mice were anesthetize by intramuscular
injection of 10 .mu.l of Zoletil 50 (50 mg/kg; Virbac Laboratories,
Carros, France), and PE-08 was cathetered in the mice (about 20-25
g of weight). Then, 200 .mu.l, A 10 mg/mL of a contrast agent with
Gd.sub.2O.sub.3 was injected from the femoral artery into the
late-stage tumor (16-day) of the mice through the above PE-08
catheters (BB31695, Scientific Commodities, Inc.: 0.2 mm, O.D.:
0.36 mm), wherein the injection velocity of each group of the
contrast agent was 1 .mu.l/s. During the imaging process, mice were
anesthetized under 1% isoflurene in oxygen. X-ray images were taken
after 1 minute from injection of the contrast agent from the
femoral artery into the mice, and the exposure time was 100
milliseconds.
[0024] Pictures of the mice sample with or without the X-ray
irradiation were taken from some examples. In comparison to the
imaging without irradiation, imaging with irradiation revealed
orange spots at the tumor sites. The orange spots from the leg
indicated Gd.sub.2O.sub.3 nanoparticles deposited at a tumor site
via intra-arterial injection.
[0025] While the invention has been described by way of example and
in terms of the preferred embodiments, it is to be understood that
the invention is not limited to the disclosed embodiments. To the
contrary, it is intended to cover various modifications and similar
arrangements (as would be apparent to those skilled in the art).
Therefore, the scope of the appended claims should be accorded the
broadest interpretation so as to encompass all such modifications
and similar arrangements.
* * * * *